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Recombinant yeast for production of the pain receptor modulator nonivamide from vanillin

Muratovska, Nina LU and Carlquist, Magnus LU (2023) In Frontiers in Chemical Engineering 4.
Abstract
We report on the development of a method based on recombinant yeast Saccharomyces cerevisiae to produce nonivamide, a capsaicinoid and potent agonist of the pain receptor TRPV1. Nonivamide was produced in a two-step batch process where yeast was i) grown aerobically on glucose and ii) used to produce nonivamide from vanillin and non-anoic acid by bioconversion. The yeast was engineered to express multiple copies of an amine transaminase from Chromobacterium violaceum (CvTA), along with an NADH-dependent alanine dehydrogenase from Bacillus subtilis (BsAlaDH) to enable efficient reductive amination of vanillin. Oxygen-limited conditions and the use of ethanol as a co-substrate to regenerate NADH were... (More)
We report on the development of a method based on recombinant yeast Saccharomyces cerevisiae to produce nonivamide, a capsaicinoid and potent agonist of the pain receptor TRPV1. Nonivamide was produced in a two-step batch process where yeast was i) grown aerobically on glucose and ii) used to produce nonivamide from vanillin and non-anoic acid by bioconversion. The yeast was engineered to express multiple copies of an amine transaminase from Chromobacterium violaceum (CvTA), along with an NADH-dependent alanine dehydrogenase from Bacillus subtilis (BsAlaDH) to enable efficient reductive amination of vanillin. Oxygen-limited conditions and the use of ethanol as a co-substrate to regenerate NADH were identified to favour amination over the formation of the by-products vanillic alcohol and vanillic acid. The native alcohol dehydrogenase ADH6 was deleted to further reduce the formation of vanillic alcohol. A two-enzyme system consisting of an N-acyltransferase from Capsicum annuum (CaAT), and a CoA ligase from Sphingomonas sp. Ibu-2 (IpfF) was co-expressed to produce the amide. This study provides proof of concept for yeast-based production of non-ivamide by combined transamination and amidation of vanillin. (Less)
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author
and
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Frontiers in Chemical Engineering
volume
4
article number
1097215
pages
10 pages
publisher
Frontiers Media S. A.
external identifiers
  • scopus:85153596766
ISSN
2673-2718
DOI
10.3389/fceng.2022.1097215
language
English
LU publication?
yes
id
82241912-bf7a-4968-87ac-62a569e01da4
date added to LUP
2023-02-03 13:53:46
date last changed
2025-04-04 15:16:47
@article{82241912-bf7a-4968-87ac-62a569e01da4,
  abstract     = {{We report on the development of a method based on recombinant yeast <em>Saccharomyces cerevisiae</em> to produce nonivamide, a capsaicinoid and potent agonist of the pain receptor TRPV1. Nonivamide was produced in a two-step batch process where yeast was i) grown aerobically on glucose and ii) used to produce nonivamide from vanillin and non-anoic acid by bioconversion. The yeast was engineered to express multiple copies of an amine transaminase from <em>Chromobacterium violaceum</em> (CvTA), along with an NADH-dependent alanine dehydrogenase from <em>Bacillus subtilis</em> (BsAlaDH) to enable efficient reductive amination of vanillin<em>.</em> Oxygen-limited conditions and the use of ethanol as a co-substrate to regenerate NADH were identified to favour amination over the formation of the by-products vanillic alcohol and vanillic acid. The native alcohol dehydrogenase ADH6 was deleted to further reduce the formation of vanillic alcohol. A two-enzyme system consisting of an <em>N</em>-acyltransferase from <em>Capsicum annuum</em> (CaAT), and a CoA ligase from <em>Sphingomonas</em> sp. Ibu-2 (IpfF) was co-expressed to produce the amide. This study provides proof of concept for yeast-based production of non-ivamide by combined transamination and amidation of vanillin.}},
  author       = {{Muratovska, Nina and Carlquist, Magnus}},
  issn         = {{2673-2718}},
  language     = {{eng}},
  publisher    = {{Frontiers Media S. A.}},
  series       = {{Frontiers in Chemical Engineering}},
  title        = {{Recombinant yeast for production of the pain receptor modulator nonivamide from vanillin}},
  url          = {{http://dx.doi.org/10.3389/fceng.2022.1097215}},
  doi          = {{10.3389/fceng.2022.1097215}},
  volume       = {{4}},
  year         = {{2023}},
}